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Serum-free culture medium system for culture of primary human epidermic cells

A technology of serum-free medium and epidermal cells, which is applied in the field of biomedical materials, can solve problems such as inevitable toxicity and influence on cell permeability, shorten the doubling time, promote cell division and proliferation, and meet the requirements of large-scale production and application Effect

Inactive Publication Date: 2017-09-05
GUANGDONG BOXI BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The application number is: 201510452454.7 Invented a serum-free culture system for epidermal cell culture, the system added amino acids, cholera toxin, growth-promoting factors, adenine, iron source, triiodothyronine to the basal medium acid and calcium salt, although this method can be used for the cultivation of epidermal cells, the cholera toxin added in it does affect the permeability of the cells, and it is inevitably toxic and has certain defects

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  • Serum-free culture medium system for culture of primary human epidermic cells
  • Serum-free culture medium system for culture of primary human epidermic cells

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Embodiment 1

[0024] Medium preparation: Mix 350 mL DMEM medium and 150 mL F12 medium to prepare a total of 500 mL; first add BPE at 0.01 mg / mL, mix well, then add L-glutamine 1 mM, EGF 0.05 ng / mL, Insulin 0.06mg / mL, penicillin: 10kU / mL; streptomycin: 10mg / mL, calcium chloride 0.5mM, transferrin 8μg / mL, adenine 200μM, hydrocortisone 0.5μg / mL, sodium selenite 15 μg / L; use HCI to adjust the pH to 7.2, and after constant volume, use a 0.2 μm filter membrane to filter and sterilize to obtain a serum-free human epidermal cell culture medium.

[0025] Example of use: wash the digested human epidermal cells with PBS solution; resuspend with the above serum-free human epidermal cell culture medium after centrifugation, inoculate 3×105 cells per bottle, and place in an incubator at 37°C, 5% CO2 Cultivate under the same conditions; observe the cell morphology after overnight, change the medium after 24 hours, continue to cultivate, and change the medium every 2 days thereafter. Primary human epiderm...

Embodiment 2

[0029]Medium preparation: various exogenous factors were added to commercial basal medium MCDB153. The concentration of BPE is 0.03mg / mL, after mixing, add L-glutamine 1mM, EGF 0.05ng / mL, insulin 0.05mg / mL, penicillin: 10kU / mL; streptomycin: 10mg / mL, chloride Calcium 0.5mM, ferric citrate 40 mg / L, adenine 200μM, hydrocortisone 0.8μg / mL, sodium selenite 15μg / L; adjust the pH to 7.2, and filter to sterilize with a 0.2μm filter membrane after constant volume. Serum-free human epidermal cell culture medium was obtained.

[0030] Example of use: wash the digested human epidermal cells with PBS solution; resuspend with the above serum-free human epidermal cell culture medium after centrifugation, inoculate 3×105 cells per bottle, and place in an incubator at 37°C, 5% CO2 Cultivate under the same conditions; observe the cell morphology after overnight, change the medium after 24 hours, continue to cultivate, and change the medium every 2 days thereafter. Primary human epidermal cel...

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Abstract

The invention provides a serum-free culture medium system for culture of primary human epidermic cells, relates to the technical field of biological medical materials, and discloses application of a serum-free culture medium in in-vitro culture of human epidermic cells. The serum-free culture medium system has the advantage that the cholera toxin and various anchoring factors are not added, the potential hidden hazard due to complicated components is avoided, the epidermic cells can well grow, and the requirement of scaled production and application is met. The serum-free culture medium system for culture of primary human epidermic cells comprises a basic culture medium and exogenous adding factors, wherein the basic culture medium is formed by mixing a DMEM (dulbecco's modified eagle medium) commercial culture medium and a F12 commercial culture medium according to a certain ratio, and can also adopt a commercialized cell culture medium MCDB153; the exogenous adding factors comprise L-glutamine, EGF (epidermic growth factors), BPE (bovine pituitary extract), insulin, penicillin-streptomycin, calcium chloride, transferrin, adenine, hydrocortisone, and sodium selenite.

Description

technical field [0001] The invention relates to the technical field of biomedical materials, in particular to a serum-free medium system for culturing primary human epidermal cells. Background technique [0002] Tissue-engineered skin is currently a hot spot in the field of tissue engineering. In recent years, many products have been used in clinical and cosmetic safety testing. A major problem to be solved in the research and development of tissue engineered skin is the large-scale cultivation of seed cells: epidermal cells. [0003] There are three main methods of culturing epidermal cells: trophoblast method, trophoblast-free method, and gas-liquid phase culture method; the culture medium used includes two kinds of culture medium with serum and without serum. The trophoblast originally used mouse fibroblasts (3T3) as a trophoblast, because 3T3 can secrete cytokines that promote the growth and division of epidermal cells, thereby promoting the growth of epidermal cells, b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
CPCC12N5/0625C12N2500/14C12N2500/25C12N2500/32C12N2500/40C12N2500/84C12N2500/90C12N2501/11C12N2501/39
Inventor 王丹丹卢永波李潇吴迪
Owner GUANGDONG BOXI BIO TECH CO LTD
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